Ultra Sensitive Rat Insulin ELISA (EIA) Kit

Catalog#
90060

Ultra Sensitive Rat Insulin ELISA Kit (Rat / Mouse Insulin Assay)

Low and High Range Rat Insulin Assay Using the Same Kit

Sample Size :

5µl

Samples:

Serum, plasma, or fluid

Tests:

96 wells (8 wells x 12 modules)

Reagents:

In liquid form (except standard)

Assay Range:

Low Range Assay: 0.1 - 6.4 ng/ml
Wide Range Assay: 0.1 - 12.8 ng/ml*
High Range Assay: 1 - 64 ng/ml

* Intended for screening purpose

Assay Time:

Same day procedure

Precision:

Intra-assay precision CV ≤ 10.0%
Inter-assay precision CV ≤ 10.0%

Recovery:

When rat insulin was spiked to rat serum, the recovery of rat insulin was 100% ± 10%

Specificity:

Substance	Reactivity
Rat Insulin	100%
Mouse Insulin	100%*
Hamster Insulin	Detected*
Rat C-peptide	Not detected
Rat pancreatic polypeptide	Not detected
Porcine glucagon	Not detected
Human Insulin like growth factor-I	Not detected
Human Insulin like growth factor-II	Not detected

*Can vary from lot to lot. Specific cross reactivity data is included with each kit

Typical Standard Curve:

Highlights:

Small Sample Volume: Only 5µl

High Sensitivity: 100 pg/ml using 5µl sample and 5 pg/ml using 100µl sample

Low and High Insulin Assay:Both using the same kit

Precision: Intra-assay and inter-assay precision CV ≤ 10.0%

Protocol: Same protocol for serum, plasma, or fluid assay

Effect of Hemolysis: Almost no effect of hemolysis tested up to 20mg/ml.

Summary of Assay Ultra Sensitive Rat Insulin ELISA Kit (Catalog# 90060)

Low/Wide and High Range Assay

Affix the antibody-coated microplate module on the frame

Dispense 95µl sample diluent per well

Pipette 5µl of the sample (or working rat insulin standard**) to the well

Incubate the microplate for 2 hours at 4°C.

Wash the well five times with washing buffer

Dispense 100µl anti-insulin enzyme conjugate per well

Incubate microplate for 30 minutes at room temperature

Wash each well seven times with washing buffer

Dispense 100µl enzyme substrate solution per well

Incubate microplate at room temperature while avoiding exposure to light

40 minutes- 10 minutes-
low/wide range assay high range assay

Stop the reaction by adding 100µl Enzyme reaction stop solution per well

Measure the absorbance at A_450nm and subtract A_630nm values within 30 minutes

Calculate the Rat Insulin concentration using the standard curve

*Each well should be washed with 300µl of wash buffer. Aspirate the wells completely so all

excess solution is removed.

** Preparation of standards are different for both low and high range assay. Please refer to the
product insert for more details

Crystal Chem Inc. Tel: (630) 889-9003 Fax: (630) 889-9021 E-mail: sales@crystalchem.com